title: A simple and rapid method to isolate purer M13 phage by isoelectric precipitation.
abstract: M13 virus (phage) has been extensively used in phage display technology and nanomaterial templating. Our research aimed to use M13 phage to template sulfur nanoparticles for making lithium ion batteries. Traditional methods for harvesting M13 phage from Escherichia coli employ polyethylene glycol (PEG)-based precipitation, and the yield is usually measured by plaque counting. With this method, PEG residue is present in the M13 phage pellet and is difficult to eliminate. To resolve this issue, a method based on isoelectric precipitation was introduced and tested. The isoelectric method resulted in the production of purer phage with a higher yield, compared to the traditional PEG-based method. There is no significant variation in infectivity of the phage prepared using isoelectric precipitation, and the dynamic light scattering data indirectly prove that the phage structure is not damaged by pH adjustment. To maximize phage production, a dry-weight yield curve of M13 phage for various culture times was produced. The yield curve is proportional to the growth curve of E. coli. On a 200-mL culture scale, 0.2 g L(-1) M13 phage (dry-weight) was produced by the isoelectric precipitation method.
authors.affiliation: Department of Chemical Engineering, University of Waterloo, 200 University Avenue West, Waterloo, ON, N2L 3G1, Canada. dongdexian@yahoo.ca
authors.forename: Dexian
authors.initials: D
authors.lastname: Dong
authors.forename: Sanjana
authors.initials: S
authors.lastname: Sutaria
authors.forename: Je Yeol
authors.initials: JY
authors.lastname: Hwangbo
authors.forename: P
authors.initials: P
authors.lastname: Chen
mesh_terms: D017104:Bacteriophage M13 / Q000737:chemistry* / Q000302:isolation & purification / Q000502:physiology; D004926:Escherichia coli / Q000821:virology; D005590:Fractional Precipitation / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point